Preparation of derivatives of 3(alpha)-hydroxycholanic acid



Patented Feb. 13, 1951 *JQPRJEPARATION OF DERIVATIVES 01 3 (oi-HYDROXYCHOLANIC ACID Edward C. Kendall, Rochester, Minn., assignor to Research-Corporation, New York, N. Y., a. corporation of New York No Drawing. Application March 8: 1946, Serial No. 653,176

1 This invention relates to the production of 3,9- epoxy steroid compounds of gthe formula CH R 19 l in which -a:yrepresents a member of the group consisting of and R represents hydrogen or a monovalent group.

In the above formula the hydrogens attached Desoxycholic acid ()GHCOO a (a) Methyl-3 u) -acetoxy-12-hydroxycholanate 8Claims. (o1.zoo-missil 5 to the carbons in positions; 1 to 17 are omitted. All substituents shown at the 3-position, that is hydro igyl, acetoxyl, benzoityl, etc. are in the 3(a)position as is'the 'hydroxyl group in desoxychloic acid. 1

An object of the invention is {the production of the 3,9-epoxy steroid compounds from 301L12- substituted steroids, as intermediates in the production of C's-substituted steroids with a keto or hydroXylgroup atCm The invention will be illustrated with reference to the preparation of methyl-3,9-epoxy-11- ketocholanate, a compound of the above formula, in which the :cy position is occupied by the 5 group m and in which position R, isvoccupied by the group An outline of the process for the preparation of this compound is shown by the following flow sheet:

Methyl desoxycholate Methyl-3 (a) -acetoxy-12-ketone cholanate 3 (a) -hydroxy-12-ketocholanic acid (a) H O- (9) Methyl-3(a)-hydwiry-12-chloro-A -cholenate Methyl-3,9-epoxy-11,12-dlbromocholanate Methyl-3,9-epoxy-l1-ketocholanate Ark J Methyl-3 (a) -hydroxy-12-methoxy- A -cholenate C H C-C-C-O CH3 CH3 Methyl-3,9-epoxy-11-keto-12-bromo cholanate 76 A crop of about 300 g. of methyl ester is obtained.

The 'flltrateis evaporated to about 300 cc. 1 On filtration'about 125 g. of themethyl ester is obtained. Further evaporation and dilution with water yields a third crop; which after recrystallization from methanol weights about 20 g. Total yield'is 440450 g. The'crystals separate from the solution combined with 0.5 mole of methanol per mole" of ester. If the desoxycholic acid is pure the yieldfis almost quantitative.

Preparation 'of methyl-3(a)'-acetoa:y-12-hydro'rycholanate" (3) ."422 g. (1' mole) of the methyl desoxycholate are suspendedin 1400 'cc."of acetic acid and 35 cc. "of 1.7 N hydrochloric acid in glacial acetic acid are added. The volume is made to 2000 co. with glacial acetic acid. The concentration'of Hpl will be 0.03 N. The solutlon is allowed to stand at 17 C. for 6 days. The methyl-3(a) -acetoxy-l2-hydroxycholanate is not separated but is used in solution for preparation of the*12-ketone derivative.

Preparation of methyl-3(a) -acetoay-lz-keiocholanate (4) .The solution of methyl-3(a) acetoxy-12-hydroxycholanate is maintained below 20 and is oxidized with 80 g. (120% of theory) of 0103 dissolved in 80cc. of water+200 cc. acetic acid. The resulting mixture is allowed to stand at 20 C. for'l hour. Four liters of water are added, the precipitated crystalline material is filtered from the solution and washed with water. The precipitate is dissolved in 2 liters of benzene and is stirred three times with 2 liters of water which contains '500 cc. of N HCl by meansof a mechanical stirrer. The benzene solution is separatedfrom the water and dried with sodium sulfate. The benzene is removed under reduced pressure. Preparation of 3(a)-h2/dTO:I2y-12-7Cet0chOla1LiC acid (5) .-800-cc. of methanol are added to the methyl-3(a) -acetoxy-lz-ketocholanate and the solution is boiled. 1000 cc. of 5N sodium hydroxide are added slowly. The methanol is'boiled off and water, toa volume of 3 liters, is added. This is allowed to cool to room temperature overnight. It is then filtered and the precipitate is washed with N sodium hydroxide. The precipitate is dissolved in-3 liters of warm water and is filtered froin suspended impurities (Cr(OI-I)s-llint). The solution is acidified with hydrochloric acid and the precipitated acid is filtered, washed by resuspens'ion in water andagain filtered. The residue dried at 110. The yield is about375g. (95-96%). M; P. 162-164"; I I

Preparation of 3(a) hydromy-lz keto A cholemc acid (6).--The residuebbtaine'dfzom preparation (4) is dissolved in 3600cc; of chlorobenzene and900 cc. of acetic acid, 5 cc. of 1 Nhydrogen chloride dissolved in acetic acid and 167 g. of'selenium dioxide areadded and the solution is boiled under a reflux condenser for 48 hours. The selenium which separates is removed byfiltrati'on and organic selenium compounds are oxidized bythe addition of g. of-chromic'acid in 200cc; ofwater. Thetwo phase system is vigorously stirred for 2 hours. The mixture is filtered and the 'chlorobenze'ne solution is washed with water to remove chromiumsalts and acetic acid."The chloro-benzene is removed'under re-' duced pressure and the residueis dissolved in methanol, 110 cc. of 18 N sodium hydroxide are added and the solution is allowed to stand a few hours tohydrolyze the methyl ester and give the carboxyr'group and to remove the acetyl group atposition 3. The methanol is removed under reduced pressure, water is added to dissolve the sodium salt the 19:1: tmpm n-F mnf'hu'nnl nvu; 1m.

mov d'and' the solution o'fffth sodiiiinfls'alt f the. acid is addedtoa flaskwliiclfccntains 3 lltersf or boiling water throughwhich a rapid streia'ni of steamf passed: 'A 'fewcirystals or 3 (a)I hydroxyl2-keto A -fch0lenic acid and somewhat, more than 12 moles of hydrochloric] acid" are added to the flask. "Thelsodiiim saltf'o'i the. cholei'iic acid is added s'lowly'untiljthe materialseparatcs in crystalline form. The alkalinelfsolution can? then be added rapi'dly'jfthe. cholenic, acid separates' as a voluminousfprecipitate." The spiuu'q'nis filtered not aiid'well Wia ueawit water The acdiis dried, at room temperature 'or slightlyialoove. nee traces of mineralfacidmaY cause'decoirfipos n- The cillde acid is dissolved in hot .80. "pfeid acetone, 20 percehflwa'teri'ri'the proportion of. 50' gm. of 'acid to of the acetone-water mixture, After the cfrude'facid has'fdissjolved the s iuuen" isplaced ice'f'bath for th separatiiiii'of the cholenicacid. suecessive'cropfsmay beseparated by concentrationf of themothe'r liq or. P; 177F178", yie1d90+,percent; In theab'ove process, th .3.,( a)'.-.jhydroxyl group may be protected by an aliphaticlonarorri'atic acyl. group whichiiis stable ,tO chromic acid, For example," the benzoyl group" may be used instead'of the acetyl group. The half ester of sue: cinic facid 6r of phthalic acid'also \l'vbuld be satis-i factory. p Preparation of 3 (d) ,12 dihydrohiy zfi if cholenic acid (7') 5-301)" hydroxyT- I21{eto A -cho1enic acidlGi is estrified iifi'methanol 'which contains 0. l N hydrochloric acid as'dfescribed for thepreparation of methyl "desoxycholat wi. 120.6, g; (0.3 mole) of the ester, are dissolved ina mixture of 300 mm ethanolfand 300 cc. oi acetic acid. 1.0g. f Adams platinum oxide is added and the fiaskli's shakenflin an atmosphere of hydrogen until 'one'molar equivalent has been absorbed'at which'time utilization of hydrogen'almost ceases. The ethanol and acetic acid arelremo ved under reduced pressure and the residue is dissolved in benzene or other organic solvent'immiscible with water. The. lasttraces of acetic acid are extracted with water and'theTbenzene is removed under reduced pressure; Thev residueis crude methyl 3(a).12dihydroxy-A l cholenate. J The 3,112 dia cetyl derivative of thislcrudefprpd uctis prepared in ac'etic anhydrideiand pyridine; This pomp'ound'is crystallized from "methanol and when so purified the methyl ester-and'the acetyl groups are removed with sodium hydroxide in methanol to 'g iv fi) ,12-dihydroXy-A -0h01e1iic acid fl). p 7 I For preparation of (8)the residueof crude methyl" ester is used; v f 1 er'epama n' 6f methyl-3 (a) -hyclro:cii-12fmethoasy-A -chdlenate 8) .-'To the residue from the preceding preparation 500 cc. of methanol, 50 cc." of"con'centrated aqueoushydro'chloric acid and-58 orwater are" added. The solution is allowed to stand at 5. Crystalsslowly separate and several successive crops may be secured. M. P. 158- 1'60.' [a]D=+ (1% in methanol). The yield is 90-} percent.

Preparation of methyl-3 (a) hydrory-IZ-chloroA =Cholenate (ML-83.6 g. (0.2l11018) of the preceding'preparatioii are dissolved 'in 40o cc. of alcohol-free chloroform in a round bottom flask. The solution is cooled to 5 in an ice bath and is vigorously stirred with 200cc. of concentrated aqueous hydrochlorica'cid. Thechlor'oform is separated and is stirred 1 twice more with 200 cc. of concentrated hydrochloric acid. The stirring chloro compound can be separated in crystalline form from the chloroform; however, for the purposes of the preparation, the subsequent step is carried out immediately on the chloroform solution. I a

Preparation of methyl 3,9-epoxy-A -cholenate (10).The chloroform solution from the preceding step is placed in a 3 or 5 liter flask with 1000 cc. of'water which contains 100 g. of sodium bicarbonate and the contents of the flask are stirred mechanically for -18 hours. During this treatment hydrogen chloride is removed from the-molecule'and the cyclic ether 3,9 is formed with a shift in the double bond from position 9,11 to 11,12. The resulting 3,9-epoxy derivative of methyl-'A -cholenate can be separated in crystalline form but for the purposes of the preparation it is left in the solution of chloroform. The chloroform is concentrated to 250 cc. The yield is almost quantitative.

Preparation of the free acid (compound 10A) from the methyl ester (compound 10).8 gm. of compound 10 are dissolved in 100 cc. of methanol with cc. of 5 N sodium hydroxide. The methanol is evaporated under reduced pressure, the residue is diluted with 80 cc. of water and kept at 0 for some hours. A crystalline sodium salt separates and is washed with ice cold 1 N sodium hydroxide solution. The sodium salt is emulsified with a mixture of 100 cc. of water and 100 cc. of chloroform and acidified with 50 cc. of acetic acid. The chloroform is washed several times with water and evaporated to dryness. The mixture is dissolved in hot 95 per cent ethanol. Crystals separate after the solution is cooled to 0". M. P. is 157-158". [a]D=51.6 (1% in methanol).

Preparation of methyl 3,9-ep0my-1L12-dibromocholanate 11).--The chloroform solution secured from preparation (10) is placed in a 500 cc. round bottom flask in the neck of which is a three-holed rubber stopper. Through one hole in the'stopper is, placed a short L glass tube to which is attached a short tube containing calcium chloridev properly protected with a plug of cotton. The stemof a 250 cc. separatory funnel is placed in the second hole of the stopper. Through the third hole in the rubber stopper one end of a long glas U tube is placed so that it will extend 'to the bottom of the chloroform solution. The other end of the lon glass U tube passes through a two-hole rubber stopper and terminates in a 1 liter round bottom glass flask. The remaining hole in therubber stopper of the 1 liter flask is attached to another calcium chloride tube which is connected with a source of vacuum. In the 1 liter flask 350; cc. of dry alcohol-free chloroform and 20.6 cc. of bromine are placed. The two flasks are placed in two large beakers'which are protected by insulation so that water does not condense on the outside surfaces. Solid carbon dioxide is placed in the two beakers and acetone is poured on it. In this way the temperature of the chloroform solution may be reduced to 78. After the flasks havebeen cooled to 40 to the chloroform solution containing the derivative of cholenic acid is drawn over by application of vacuum into the 1 liter flask which contains the chloroform and bromine. This step is carried out slowly and the total. volume of chloroform is added in three portions, approximately one-third at a time. A short interval is allowed to lapse between addition to permit the temperature to be maintained at approximately 50. A small amount of heat is produced by adding bromine to through the acetone.

8 i the double bond. This is indicated by an increased rate of bubbling of the carbon dioxide 50 cc. of chloroform are then added through the separatory funnel of the 500 cc. flask and after it is cooled to 50 it is drawn over into the 1 liter flask containing the bromine. This'is followed by two additional 50 cc. portions of chloroform. The bromination is allowed to proceed for 30 minutes. The apparatus is then disconnected and the 1 liter flask which contains the react-ion mixture is allowed to warm up until the chloroform which has frozen melts- The chloroform is then poured into a 4 liter beaker which contains 1700 cc. of water in which are dissolved 35 g. of sodium sulflte and 35 g. of sodium bicarbonate. The beaker isplaced under a mechanical stirrer and is stirred for 30 minutes. The chloroform solution is separated andwashed with distilled water. 250 cc. of methanol are then added and the mixture of chloroform and methanol is concentrated to about 225 cc. under reduced pressure or until crystals or a turbidity develops. If the solution becomes turbid, it is removed, diluted with 40 cc. of methanol and a few crystals of the dibromo derivative are addedL This results in prompt formation of crystals through the solution. 460 cc. of methanol are added andthe solution is allowed to stand at room temperature for /4 l;1our. It is then filtered and the separated crystals are washed with two 100 cc. portions of cold'meth anol. These crystals usually melt at 1415-143" and weigh 65-70 g. to per cent of theory). The dibromo compound is purified by solution in a small volume of hot benzene to which warm methanol is added. Crystals soon separate and are filtered'off after the solution has been cooled. The pure compound melts at 143. [a]D=+52 (1% in chloroform), V The mother liquor of the crystals is seeded with the dibromo ester which melts at 123 and is concentrated under reduced pressure to about 200 cc. The crystals are washedwith three 50 cc. portions of cold methanol and dried. The yield is 20 to 25 g. The melting point is usually 100-112 These crystals are a mixture of two dibromo compounds which differ only in the spatial relationships of the bromine atoms at 011,12. One of the components melts at 143 and the other at 123. After about 0.2 mole of this mixture has been accumulated it is converted into methyl 3,9-epoxy-A -cholenate through debromination and this is rebrominated precisely as described. For the debromination 119 g. of the mixture melting at'100-112" is dissolved in 230 cc. of boilin ether and 77 g. of powdered zinc are added. 770 cc. of methanol are added slowly with stirring and. heating. The reaction mixture is heated in a 2 liter round bottom flask on the steam bath for fifteen minutes and. cooled to room temperature. The zinc is filtered from the solution, methanol and remaining traces of ether are removed under reduced pressure to a volume of 200 cc. 500 cc. of chloroform are then added and the methanol is washed from the chloroform with eight successive 1 liter portions of water. It is best to filter the chloroform and water through a pad of infusorial earth after the first addition of water to the methanol and chloroform. The zinc salts which are present precipitate in part. After filtration, the chloroform may be washed with water without difliculty. After the methanol has been removed the chloroform solution is dried with sodium sulfate, concentrated under reduced pressure to about 250 cc. and is then brominated precisely as described-.2

Preparation of methyl 3,9-epoxy-11-7ceto-12- bromocholanate (12).-109 g. of methyl 3,9- epoxy-l1,12-dibromocholanate is suspended in 2500 cc. of ll. S. P. acetone in a flask equipped for mechanical stirring; The flask is placed in a water bath the temperature of which is maintained at 25 throughout the, .reaction. While I? the suspension is being stirred 50 g. of powdered silver chronate and a solution oi 38.6 g. of chromium trioxide in 243 cc. of ,water are added in single portions. The temperature .oi the reaction mixture within the flask rises to about 30 within minutes after addition of the silver chromate and chromium trioxide. The suspension is stirred mechanically until all of the crystals zine-td emmaecemneuet eq se re o 2 H hours) .and is I allowed to stand without stirring for 15 hours at25. '77 co nt 5 ll sulfuridacid are added with stirring and the suspension,

stirred for thirty minutes; .The solution is ,,d

cantedfrom the ihurgaLnidsalts and filte red The residue is washed with two 100 t c tion. If the separation of crystals causes bumping, the crystals are remoyed and the concentration of the mother. liquor is then continued until thevolume of the solution is between 450 and 500 cc. (If the solution is concentrated to a smaller volume a gum separates and contaminates the product.) with water and dried. If the crystals are separated in two crops the first usually melts at 114- 115 and the second at about 112- 114". The total yield is from 92-94 percent. chloroform) Preparation of methyl 3,9-epowy -11-ket0- cholanate (13) .14.4 g. (0.03 mole) of methyl 3,9-epoxy-11-keto-12ebromocholanate are dis- The crystals .are filtered, washed well solved in 200 cc. of glaciall acetic acid and 15 g.

of powdered zinc are added. stirred with a mechanical stirrer for 15-20 minutes at room temperature. The zinc is removed by filtration, the acetic acid is removed under reduced pressure, 100 cc. of benzene and 200 cc.

of water are added and the benzene is then washed with water to remove acetic acid and zinc salts. The benzene is removed under reduced pressure and the methyl 3,9-epoxy-1lketocholanate is crystallized from dilute methanol. M. P. 77-88". [a] D=+88 (1%in chloroform).

All of. theealzzove lcompoundsw erivedir desoxycholic acid, contain the sid chain thereof in 17-positi0n. Compounds containing other substituents in the l'l-position may be made, e. g.

by the use ofother 3( 12-sub stituted steroid compounds or by the removal or shortening of the 17-side chain at the beginning or. at some The solution is intermediate stage or at th end of the synthesis of compound 13 outlined above;-

Degradation of the side chain attached to the carbon in 17-positionto produce compounds in which the R of the general formula given at the beginning of this specification. represents:

in which Z stands for hydrogen-or an alkyl group is illustrated by the following.

he side h n m e esred st p r.9 1 carbon at time byametlrod devised by Wieland unnecessary to describe it further than to state that the Grignard reaction using magnesium phenylbromide is employed to replace the ester groups and forrn. the corresponding diphenyl carbinols, -which latter are converted into the corresponding diphenylethylenes and these are oxidized to give diphenyl ketgne and the acid on the side chain with one carbon atom less than in the starting compound. It may be noted, however, that the known WielandandBarbier method is improved or gives improved yields if the Grignard reaction is carried out at a low temperature and for a longer time, e. g. at 0 C. for 24 hours instead of at high temperature for several hours. A furtherximprovement inLWieland and Barbier method. s ,ig.cer .y. tbe oxidation with chromic acid in the presence of acetic acid, chloreform and water, the presence of which latter prevents iormationgf an oxide at th double bond and favors formation of the glycol which is easily oxidized to the next lower acid. A further improvement ,in the Wieland and Barbier method is the addition; of sulfuric acid to the chromic acid used in ;the oxidation to form chromium sulfate1..from. the reduced chromium instead of chromium acetate. For example, for the oxidation of 0.1 mole of the; ethylene compound I may use a mixture of 155 co or chloroform, 100 cc. of 10 N chromic acid dissolved in a mixture of acetic acid and 15% water and cc. of 10 Nsulfuric acid inamixture of 85% acetic acida'nd 15%wa'ter. This same solution maybe used for the oxidation of the first ethylene from ch olanic acid and the second ethylene from thenor acid. Still another improvement in; the Wielandand Barbier method is the use of a solvent such'as ethyl Inorpholine to keep th intermediate prod'uct in solution during the formation of the diphenyl carbinol. This sequenceof steps, starting withcompound 12 and ending with compound 21, is showii in the following flow sheet: y

Compound 12 3.9-ePOXy-11-ketonorcholanyldiphenylcarbinol CH3 w 5; 2.

(l6) 3,9-epoxy-11-ketobisnorcholanyldiphenylethylene r 2,541,014 v V 11 v 12 CH;- CH; 7 H-C-COOH v HI-COOCH; cm aI j 6 x I i V 3,9-epoxy-11-ket no ho1 n1 acid 15 Methyl-3,9-epoxy-1l-ketobisnorchoianate In proceeding from compound 21 the degrada- 7 tion procedure is the same as above excepting H( Jg-0-i-00CH= that the oxidation is accomplished by means of I ozone, instead of chromic acid, which breaks off the side chain attached to the 2 0-carbon and 0: gives compound 24. This sequence of steps is represented by the following flow sheet:

- Compound 21 CH! CcH H "(J-(MB; v (11) 1 0H Methyl 3,9-epoxy-11-ketonorcholanate 30 (2 3) 3,9-epoxy-l1-ketomethyletiocholanyldiphenylethylene C: CH3 I Q; 9 3,9-epoxy-ll-ketopregnanyldiphenylethylene 6 1?)\ OH; I

H -CO0H 0: 3,9-epoxy-11,ZO-pregnanedione In order to convert compound 24 into the compound having the carboxyl group at position 17, N compound 24 is treated with benzaldehyde in methanol and sodium methylate. This is a, known 1 ype of reaction and yields compound 25 shown I V l v c 7 below. This benzal derivative is then treated with ozone which liberates benzaldehyde and (20) forms an aldehyde group at the 21 carbon atom 39.93amLketobisnorcholanic acid and the resulting compound upon treatment with periodic acid yields the etio acid compound ggi as follows: Compound 24 raw (11mm- A u.

3,9-epoxy-ll,20-pregnanedione-Zl-benzal 6 3,9-epoxy-1l-ketoetiocholanic acid The same treatment with benzaldehyde and sodium methylate can be applied to the 3(a)-hydroxy derivative after the-epoxy structur has been opened. This'gives 3(d") -hydroxy-11-ketoetiocholanic acid after ozonization of the 21 benzal intermediate and treatment with periodic acid. Also the side chain can be de'graded with the 3(a) -hydroxy series ofintermediates instead chromicacid in. each' case.

. group must be .acetylated before 4 oxidation with The methyl ester of compound 26"may be"preancl and mineral acid. forinstance, sulfuric acid o'f"1% concentration." v P 1 Another method for removing three carbons of the side cl fain' at CEO which is aiipncabietome 3,9 epoxy'compo'unds referred to above as well anyldiphenylethylene is as follows. This method involves the formatio'nof' a'compou'ridwith two .pared by. treating it. atboiling pointwith methdouble bonds" in the side chain in 'pdsfitions2 02 2 and- 23-24, i. e; the diene "compound, "an may start with compound 15 01 ZQWhiCh a1ready have i a double b'ond'in the side chain at t e 23-24 esince. "One method forthe introduction of the seabed double bond in the'side"chain is rotten compound 15 in an organic solvent suchasfben- 'zene, chlorobenzene, chloroform orfearbon tetrachloride 1 with selenium dioxide inth'e resence of'pyridine. The resultds 'thelntro'duction of "the hydroxyl'grou'p o'ntl'ie carbon in22"positio'n.

Theresulting ambourine? :1 when trea fwitha Another-method for thefiintroduction o'ffthe second'doublabond inthe side chain which is applicable generally to 3,9-epoxy and 3(a)-S1.1bstituted=ll keto steroid compounds isby treating the diphenylethylene"derivative, e. g. compound 29, with b1'omo"succinimide in "carbon; tetrachlor'ider'lhe reaction is catalyae'd'by exposure tolight with wave lengths in the visible spectrum as by exposing the reaction mixture to the light from an ordinary electric light bulb for about 15 minutes. The resulting compound 30 containing bromine substituted on the 22-carbon is separated from the succiri'ifnide formed, which separates in the reaction mixture asan insoluble product, and may be converted into the 'dienecompound 31 by refluxing in carbon tetrachloride or by heating with acetic acid and sodium acetate. A specific example of this procedure is as ffollows: Forty grams of 3(a) -acetoxy-ll-keto-l2' bromobisnorcholanyldiphenylethylene were heated to boiling in 400 cc. of carbon tetrachloride with 11.32 gm. of bromosuccinimide for 20 minutes over a 200 watt bulb. The solution was'cooled, filtered, carbontetrachlor-ide wasremoved under reduced pressure, 400 cc. of acetic acid and 21 gm. of sodium acetate were added and the solution was heated to boilin'g-forso minutes. The acetic acid was removed under reduced pressure and the residue was dissolvedfin benzene and washed with water. The benzenev'vas removed to small volume, acetic acid added and the last traces of benzene were taken off under reduced pressure. Crystals separatedfroin solutiori'which were filtered and purified by treatment with 150 cc. of acetone at the boiling point. :The solution was cooled in an ice bath and filtered. The crystals, M. P. 179-180, were 3(a)-acetoXy 11 keto-l2- bromo 24 diphenyI-A -clfoladiene. The action-of the'bromosuccini de-was'to introduce a bromine group at 22 andh drobromic acid was removed between 20 and "22. through the action of sodium acetate in acetic acid.

For the oxidation of "the diene (31), 5.8 gm. of the diene weredissbl'vd infi?) cc. of acetic acid and 37 cc. of chloroform (free from alcohol), 9.2 cc. of water and 16.6 cc, of 10 N chromic acid in per cent acetic acid and 15 per cent water were added to the solution cooled to 10. After fifteen minutes 16.6 cc; oflO N sulfuric acid in 85 per cent acetic acid and 15*per cent water were added and the solution was stirred mechanically one hour and forty-five minutes; the temperature was between l0-15. The solution was diluted with water and extracted with chloroform. From the chlorof'drnl the 3(a) acetoxy 12 bromo-11,20-pregnanedione was separated and crystallized from petroleum ether. M. P. 186.5-188", yield about 65 per cent.

The two methods described above for forming the diene compoundsare illustrated by the following flow sheet; h v p M v p Selenium dioxide 3(a) acetoxy-l1-keto-12-bromobisnorcholany1- (a) OAC-- l 3 a) -acetoxy-1l-keto-12-b1'omo-Z-diphenyl-A it choladiene The diene side chains of the compounds 28 and 31 may be removed by oxidation with chromic acid as described above which removes the whole side chain and forms the C ketone. The 3,9-epoxy group in the one instance and the 3()-acetoxy group, the ll-keto group and the l2-brom'ine in the other instance ,remain unwashed with water.

altered. The respective products are compounds 24 and 32:

' Compound 28 V 8.9-epoxy-ILZO-pregnandione Compound 31 o (a 0A c2) 3 (a) -acetoxy-12-bromo-11,20-pregnandione r C=O Br CH3 the more readily is the epoxy structure opened It is preferred therefore to open the epoxystruoture at the stage of compound 13 or compound 15. This process is illustrated by the following example:

Preparation of methyl 3(a)-hydroa:y-11-Icet0- cholanate (35) from methyl 3,9-epoxy-11-ketocholanate (13).-Twenty grams of methyl 3,9- epoxy-ll-ketocholanate (13) are placed in a. heavy walled glass pressure bottle. Twenty-five cc. of dry chloroform and 25 cc. of acetic anhydride are added and the flask is cooled in an acetone-Dry Ice bath to approximately -78.

Into the flask gaseous hydrobromic acid is passed until the weight of the flask is increased 100 gm. The flask is securely stoppered with a glass block and rubber ring and is placed in an ice bath. The temperature is maintained at 0 for twentyfour hours. The flask is again cooled in acetone and Dry'Ice and when the temperature is approximately 78 the stopper is removed. At the temperature of Dry Ice the pressure in the bottle is approximately atmospheric pressure; at 0 the pressure increases to about 70 pounds per square inch. The contents of the flask are immediately poured into a beaker on cracked ice. Chloroform is added and the solutionis well The chloroform is concentrated to about cc.; 500 cc. of methanol and 5 cc. of acetyl chloride are added. The hydrochloric acid liberated from the acetyl chloride in the presence of the methanol restores the methyl ass-seer 1-7 ester whi h may have been removed during treatment with hydrobromic acid. At the same time the methanolic hydrogen chloride removes the acetyl group at position 3 and forms the free hydroxyl group. Methanol is removed under reduced pressure. The residue is dissolved in chloroformor benzene and is washed with water to remove acid and methanol. Most of the solvent is removed and to the residue ligroin or petroleum ether isadded. The methyl 3(a) -hydroxy-ll-keto-l2-bromocholanate (34) crystallizes from the solvent M. P. 154. This bromo derivative is debrominated by Solution in glacial acetic acid through the addition of zinc dust. The excess zinc is filtered from solution and the acetic acid is, removed, under reduced pressure. The residue is dissolved in benzene and washed with water; the benzene is dried and taken to a small volume: The ester crystallized from llgroin. M. 1?. 100. The free acid caste-separated after hydrolysis of the methyl ester M. P. 186

Compound 13 CH3 C O O- 33) Methyl-3 (a)-acetoxy-11-keto-12-bromo-cholenate (so Methyl-3 (a)-hydtext-1i-kett-m bromo-eiitiaiiere this through the use of gaseous hydrogen bro midepasscd into a solution at such a tempera:- ture that the gas is condensed to liquid hydrobromic acid. It not essential to use acetic anhydride in the reaction. This Was done the example givenabove to prepare the; acetate ester at the B-hydroxy group as rapidly as it was opened. The purpose of the acetylation at 3 Was to diminish or block the introduction of bromine by substitution of the hydroxyl group at 3. Through the action of hydrobromic acid I have opened the epoxy structure in a chloroform solution alone without acetic anhydride. If the concentration of hydrobromic acid is too great, and if the solution is allowed to stand several days, the hydroxyl or acetyl group at 3 is replaced with bromine with formation of the 3,12-dibromo derivative.

An alternate method for the production of methyl- 3;9-epoxy-n eholenate (16) is by starting with 3(a)-hydroxy-11,12-dibrornocho lanic acid, treating it with sodium hydroxide to the production of. the 3(a) ,-1-2-dihydroxy.-A cholenic acid (7), esterifying the carboxyl group and forming the 12-methoxyl group by treating With methyl alcohol and mineral acid thereby forming mthyl 3 (a) -'hydroxy-'-l2-methoxy-A cholenate. (8), replacing the methoxyl group by chlorine by treatment with hydrochlori'ciacid thereby forming methyl-3 (a) -hydroxy-12 -chloros crieienate (9), and removingth'e hydrogenoi the 3 hydroxy group and the 12 chlorine" arid with a double bond at C11; C12.

The reaction can be carried out otherwise than by thegusev of sodiumbicarbonate as described. For examplqii the. l2-halogen compound (compound 9) is dissolved in chloroform and the chloroform solution is then Washed with water, halogen acid is removed from the chloroform as fast as it is formed and. the reaction will proceed in the case of the 12-bromo compound withgreat speed; in the case of the 12=chloro, it proceeds, more slowly. The addition of any base'suchas' sodium carbonate, sodium bicarbonate, sodium hydroxide or calcium carbonate will neutralize the acid as it is' formed in the aqueous chloroform mixture. A very satisfactory alternate procedure is to dissolve the 12- halogen compound in chloroform and add pyridine. After several hours an excellent yield of the epoxy compound is secured.

I have prepared dehydrocorticosterone from the etio acid prepared by degradation of the 1'?- side chain of compound 35 to theetio acid, and then building up thetwo carbon side chain at sition-Tf by the "method used byneichstein'icr preparation of desoxycorticosterone. 3(a)-acetoxy-11-ketoetiocholahic acid was converted vinto the acid chloride which latter was treated with diazo methane and then with potassium hydroxide to give 3(a)hydroXy-2l-diazo- V 11,20-pregnanedione; ,Acetic acid converted the C21 diazo group to the C21 acetoxy1 group. Oxida- That is,"

tion of the 3(a) -hydroxy group with chromic acid gave 21-acetoxy-3,11,20-pregnanetrione. Bromination at Grand dehydrobromination gave 21- acetoxy-A -3,11,20 pregnentrione.

; Iclaim:

1 1. As a new product a compound of the formula a in J in which It stands for a member of the groupconsisting of in which X stands for a member of the group consisting of hydrogen and the lower alkyl groups.

2. As a new product a compound as defined in claim 1 in which R represents the group CH; 0 HJ-CHr-CHi--OX 3. As a new product a compound as defined in claim 1 in which R represents the group on, o HJ'},OH:EIJOX 4. As a new product a compound as defined in claim 1 in which R represents the group 5. As a new product a compound as defined in claim 1 in which R represents the group i l-ox l 6. Process for the production of a compound of the formula in which R stand for a member of the group consisting of CH1 0 O H+ -ox; and i l-ox in'which X stands for a member of the group consisting of hydrogen and the lower alkyl groups which comprises removing halogen acid from a compound of the formula by contacting a solution thereof with an aqueous solution of a base. I K I \l I 7. Process for the production of a compound of theformula c oei-oona Hal Ill

by contacting a solution thereof with an aqueous solution of a base. v

8. Process as defined inclaim '7, in which the removal of halogen'a ci'd is efiected in the presence of chloroform and an aqeous solution of an alkali.

EDWARD C. KENDALL.

7 REFERENCES CITED The following references are of record in the file of this patent: UNITED' STATES PATENTS Name Date Wallis July 12, 1938 Nov. 25, 1941 Mar'.2, 1943 Wallis Feb. 8, 1944 FOREIGN PA'IENTS Country Date :1 Great Britain Mar. 25, 1942 OTHER REFERENCES v Ziegler: Annalen, 551, pp. 80-119 (1912);.

Number Number 

1. AS A NEW PRODUCT A COMPOUND OF THE FORMULA 